Heteroduplex Mobility Assay (HMA) Using MultiNA

Introduction

The arrival of genome editing tools that make use of TAL (transcription activator-like) effector nuclease and CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins 9) has enabled genes to be specifically destroyed and introduced with respect to target genes. These tools are rapidly becoming popular because they can now be applied to organisms, such as microorganisms, animals, and plants, for which genetic modification has proved difficult. The evaluation of whether or not there are mutations at target regions can be done by either directly analyzing genetic sequences or by utilizing enzymes that recognize and cut mismatches in duplexes. However, these methods are expensive and laborious. Heteroduplex mobility assay (HMA), on the other hand, is a method that can be performed simply, rapidly, and inexpensively (Fig. 1). In normal electrophoresis, DNA is a homoduplex of perfectly complementary strands and mobility depends on molecular weight (size). However, in DNA where either of the two strands has a mutation, the mismatched section does not form a complementary strand and as such becomes heteroduplex DNA. Mismatched sections of heteroduplex DNA have a different spatial structure to that of homoduplex DNA. For this reason, the mobility of heteroduplex DNA in electrophoresis has a tendency to be slower. By utilizing this phenomenon, HMA can determine the presence or absence of mutations and judge genotypes through electrophoresis. This article introduces an example of analysis where HMA detection with model DNA is performed using the MCE-202 MultiNA microchip electrophoresis system for DNA/RNA analysis.

November 30, 2017 GMT